Molecular and phenotypic characteristics of T1 transgenic yellow cosmos (Cosmos sulphureus) carrying neomycin phosphotransferase II gene

##plugins.themes.bootstrap3.article.sidebar##

PDF
DOI https://doi.org/10.13057/biodiv/d231202
Issue
Vol. 23 No. 12 (2022)
Section
Articles
Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

##plugins.themes.bootstrap3.article.main##

MUHAMMAD BURHANUDDIN IRSYADI
Study Program of Plant Breeding, Faculty of Agriculture, Universitas Gadjah Mada. Jl. Flora, Bulaksumur, Sleman 55281, Yogyakarta, Indonesia
WIDHI DYAH SAWITRI
Study Program of Plant Breeding, Faculty of Agriculture, Universitas Gadjah Mada. Jl. Flora, Bulaksumur, Sleman 55281, Yogyakarta, Indonesia
AZIZ PURWANTORO ♥
Study Program of Plant Breeding, Faculty of Agriculture, Universitas Gadjah Mada. Jl. Flora, Bulaksumur, Sleman 55281, Yogyakarta, Indonesia

Abstract

Abstract. Irsyadi MB, Sawitri WD, Purwantoro A. 2022. Molecular and phenotypic characteristics of T1 transgenic yellow cosmos (Cosmos sulphureus) carrying neomycin phosphotransferase II gene. Biodiversitas 23: 6097-6105. Yellow cosmos (Cosmos sulphureus) is a tropical ornamental flower that contains secondary metabolites useful fornatural pesticide application. Plant genetic transformation is one genetic engineering method used to increase secondary metabolite accumulation. However, published reports on this issue have yet to be made available. This is the first report of genetic transformation in transgenic yellow cosmos using neomycin phosphotransferase II (nptII) encoding gene. This study aimed to determine the efficiency transformation and phenotypic character of the T1 yellow cosmos transgenic nptII. This genetic transformation was carried out by the floral dip method mediated with Agrobacterium tumefaciens and characterized based on quantitative and qualitative observations, as well as a cluster analysis of transgenic plants. The T1 transgenic yellow cosmos was confirmed using PCR with a transformation efficiency of 73.33% based on the total plants resistant to Kanamycin and 10.57% based on the total seeds transformed. The presence of the nptII encoding gene was shown in transgenic plant samples with a size of 550 bp. In general, the introduction of the nptII gene generated no novel traits except the resistance to the kanamycin antibiotic. However, there was a decrease and increase in the number of stomata and the size of stomata in transgenic plants, respectively. In addition, there was a change in the type of ray floret to a mixture of ligulate and tubular, indicating that the nptII gene affected the phenotype of yellow cosmos. The result revealed that the nptII gene was inserted fairly randomly into the plant genome.

##plugins.themes.bootstrap3.article.details##

References
Aftab, N., K. Saleem, A. H. A. Khan, T. A. Butt, C. R. Mirza, J. Hussain, G. Farooq, A. Tahir, S. Yousaf, M. I. Zafar, I. Nawaz, and M. Iqbal. 2021. “Cosmos Sulphureus Cav. Is More Tolerant to Lead than Copper and Chromium in Hydroponics System.” International Journal of Environmental Science and Technology 18(8):2325–34. doi: 10.1007/s13762-020-02981-w.
Alvarez, Maria alejandra. 2015. Genetic Transformation. Rijeka: Janeza Trdine.
Bakhsh, Allah, Emine Anayol, and Sancar Fatih Ozcan. 2014. “Comparison of Transformation Efficiency of Five Agrobacterium Tumefaciens Strains in Nicotiana Tabacum L.” Emirates Journal of Food and Agriculture 26(3):259–64. doi: 10.9755/ejfa.v26i3.16437.
Chen, Ren, Mayumi Gyokusen, Yoshihisa Nakazawa, Yinquan Su, and Koichiro Gyokusen. 2010. “Establishment of an Agrobacterium-Mediated Transformation System for Periploca Sepium Bunge.” Plant Biotechnology 27(2):173–81. doi: 10.5511/plantbiotechnology.27.173.
Chetty, V. J., N. Ceballos, D. Garcia, J. Narváez-Vásquez, W. Lopez, and M. L. Orozco-Cárdenas. 2013. “Evaluation of Four Agrobacterium Tumefaciens Strains for the Genetic Transformation of Tomato (Solanum Lycopersicum L.) Cultivar Micro-Tom.” Plant Cell Reports 32(2):239–47. doi: 10.1007/s00299-012-1358-1.
Curtis, I. S., J. B. Power, P. Hedden, A. Phillpis, K. C. Lowe, D. A. Ward, and M. R. Davey. 2000. “Transformation and Characterization of Transgenic Plants of Solanum Dulcamara L.ÐIncidence of Transgene Silencing.” Annals of Botany 86:63–71. doi: :10.1006/anbo.2000.1159.
Davey, M. R., J. R. Sonej, M. N. Rao, S. Kourmpetti, and A. Bhattacharya. 2010. “Generation of Development of Transgenic Crop Plants: An Overview.” Transgenic Crop Plants 1(1):1–30.
E C Ulian, J. M. Magill, C. W. Magill, and R. H. Smith. 1996. “DNA Methylation and Expression of NPT II in Transgenic Petunias and Progeny.” Theor Appl Genet 92:976–81.
Fatmawati, Yeni, agus budi Setiawan, Aziz Purwantoro, dyah weny Respatie, and chee how Teo. 2021. “Analysis of Genetic Variability in F2 Interspecific Hybrids of Mung Bean (Vigna Radiata) Using Inter-Retrotransposon Amplified Polymorphism Marker System.” Biodiversitas 22(11):4880–89. doi: 10.13057/biodiv/d221121.
Fibriani, Suwinda, Inyana Agustien, Widhi Dyah Sawitri, and Bambang Sugiharto. 2019. “Genetic Transformation and Expression of Sucrose Phosphate Synthase Mutant in Tomato Plant.” J. Bioteknol Biosains Indones 6(1):134–43.
Franks, Tricia, Ding Gang He, and Mark Thomas. 1998. “Regeneration of Transgenic Vitis Vinifera L. Sultana Plants: Genotypic and Phenotypic Analysis.” Molecular Breeding 4:321–33.
Freiman, Aviad, Lyudmila Shlizerman, Sara Golobovitch, Zeev Yablovitz, Raia Korchinsky, Yuval Cohen, Alon Samach, Elisabeth Chevreau, Pierre Marie Le Roux, Andrea Patocchi, and Moshe A. Flaishman. 2012. “Development of a Transgenic Early Flowering Pear (Pyrus Communis L.) Genotype by RNAi Silencing of PcTFL1-1 and PcTFL1-2.” Planta 235(6):1239–51. doi: 10.1007/s00425-011-1571-0.
Imaniar, Rizki, Latifah, and Warlan Sugiyo. 2013. “Ekstraksi Dan Karakterisas Senyawa Bioaktif Dalam Daun Kenikir Sebagai Bahan Bioimsektisida Alami.” Indonesian Journal of Chemical Science 2(1):51–55.
Irsyadi, Muhammad Burhanuddin. 2022. “Penyisipan Gen SoSPS1 Secara Floral Dip Melalui Agrobacterium Tumefaciens Pada Tanaman Kosmos (Cosmos Sulphureus Cav.).” [Thesis]. Universitas Gadjah Mada, Yogyakarta. [Indonesia]..
Isda, Maya Novaliza. 2012. “Optimization of Concentration Kanamycin in the Soybean Explants (Glycine Max L.) to Transformation TcPIN Gene.” in Semirata PTN Barat. Universitas Negeri Medan, Medan, 11-13 Mey 2012. [Indonesia].
Kishimoto, Sanae, and Akemi Ohmiya. 2006. “Regulation of Carotenoid Biosynthesis in Petals and Leavesof Chrysanthemum (Chrysanthemum Morifolium).” Physiologia Plantarum 128:436–47. doi: 10.1111/j.1399-3054.2006.00761.x.
Kuluev, B. R., A. B. Knyazev, Ya. P. Lebedev, and A. V. Chemeris. 2012. “Morphological and Physiological Characteristics of Transgenic Tobacco Plants Expressing Expansin Genes: AtEXP10 from Arabidopsis and PnEXPA1 from Poplar.” Russian Journal of Plant Physiology 59(1):108–17. doi: 10.1134/S1021443712010128.
Learn-Genetic. 2018. “The Genetics of Flower Color.” Https://Learn.Genetics.Utah.Edu/Content/Flowers/Genetics. Retrieved (https://learn.genetics.utah.edu/content/flowers/genetics).
Liu, Yanbin, Chong Mei John Koh, Longhua Sun, Mya Myintzu Hlaing, Minge Du, Ni Peng, and Lianghui Ji. 2013. “Characterization of Glyceraldehyde-3-Phosphate Dehydrogenase Gene RtGPD1 and Development of Genetic Transformation Method by Dominant Selection in Oleaginous Yeast Rhodosporidium Toruloides.” Applied Microbiology and Biotechnology 97(2):719–29. doi: 10.1007/s00253-012-4223-9.
Lukmanasari, Putri, Aziz Purwantoro, Rudi Hari Murti, and Zulkifli. 2020. “Similarity Level of Nepenthes Spp. Based on the Qualitative Characters.” Ilmu Pertanian (Agricultural Science) 5(1):140–49. doi: doi.org/10.22146/ipas.55728.
Lynch, P. T., J. .. Jones, N. .. W. Blackhall, M. .. R. Davey, J. B. Power, E. .. C. Cocking, M. R. Nelson, D. M. Bigelow, T. V. Orum, C. E. Orth, and W. Schuh. 1995. “The Phenotypic Characterisation of R2 Generation Transgenic Rice Plants under Field and Glasshouse Conditions.” Euphytica 85:395–401.
Marenkova, T. V., V. V. Kuznetsov, and E. V. Deineko. 2021. “Features of Expression of Foreign Genes in Complex Insertions in Transgenic Tobacco Plants with a Mosaic Pattern of NptII Gene Expression.” Russian Journal of Genetics 57(3):319–28. doi: 10.1134/S1022795421030108.
Marenkova, T. V., Y. V. Sidorchuk, V. V. Kusnetsov, and E. V. Deineko. 2020. “Effect of Changes in Genome Ploidy on the Mosaic Character of NptII Gene Expression in Epialleles of the Transgenic Tobacco Line Nu21.” Russian Journal of Genetics 56(2):204–12. doi: 10.1134/S1022795420020088.
Messmer, M. M., A. E. Melchinger, J. Boppenmaier, R. G. Herrmann, and E. Brunklaus-Jung. 1992. “RFLP Analyses of Early-Maturing European Maize Germ Plasm - I. Genetic Diversity among Flint and Dent Inbreds.” Theoretical and Applied Genetics 83(8):1003–12. doi: 10.1007/BF00232964.
Mustofa, Z., I. M. Budiarsa, and G. B. N. Samdas. 2013. “Variasi Genetik Jagung (Zea Mays L.) Berdasarkan Karakter Fenotipik Tongkol Jagung Yang Dibudidaya Di Desa Jono Oge.” Jurnal Ilmiah Pendidikan Biologi 1:33–41.
Peña, Pamela A., Truyen Quach, Shirley Sato, Zhengxiang Ge, Natalya Nersesian, Ismail M. Dweikat, Madhavan Soundararajan, and Tom Clemente. 2017. “Molecular and Phenotypic Characterization of Transgenic Wheat and Sorghum Events Expressing the Barley Alanine Aminotransferase.” Planta 246(6):1097–1107. doi: 10.1007/s00425-017-2753-1.
Pons, Elsa, Josep E. Peris, and Leandro Peña. 2012. “Field Performance of Transgenic Citrus Trees: Assessment of the Long-Term Expression of UidA and NptII Transgenes and Its Impact on Relevant Agronomic and Phenotypic Characteristics.” BMC Biotechnology 12(41):1–15. doi: 10.1186/1472-6750-12-41.
Rashid, Abdur. 2017. “Comparison of a Kanamycin versus Hygromycin Resistance Gene in Transgenic Plant Selection of Arabidopsis Thaliana L.” Adv Cell Sci Tissue Cul 1(1):1–2.
Respatie, Dyah Weny, Prapto Yudono, Aziz Purwantoro, and Y. Andi Trisyono. 2019. “The Potential of Cosmos Sulphureus Flower Extract as a Bioherbicide for Cyperus Rotundus.” Biodiversitas 20(12):3568–74. doi: 10.13057/biodiv/d201215.
Rod-In, W. Sujipuli, and K. Ratanasut. 2014. “The Floral-Dip Method for Rice (Oryza Sativa) Transformation.” Journal of Agricultural Technology 10(2):467–74.
Sarkar, Sayantika, and Sumita Jha. 2021. “Effects Associated with Insertion of Rol Genes on Morphogenic Potential in Explants Derived from Transgenic Bacopa Monnieri (L.) Wettst.” Plant Cell, Tissue and Organ Culture 146(3):541–52. doi: 10.1007/s11240-021-02092-5.
Schuh, W., M. R. Nelson, D. M. Bigelow, T. V. Orum, C. E. Orth, P. T. Lynch, P. S. Eyles, N. W. Blackhall, J. Jones, E. C. Cocking, and M. R. Davey. 1993. “The Phenotypic Characterisation of R 2 Generation Transgenic Rice Plants under Field Conditions.” Plant Science 83:69–79.
Setiawan, AB, CH Teo, S. Kikuchi, H. Sassa, K. Kato, and T. Koba. 2020. “Centromeres of Cucumis MeloL. Comprise Cmcent and Two Novel Repeats, CmSat162 and CmSat189.” PLoS One 15:e0227578. doi: 10.1371/journal.pone.0227578.
Sharon, Kadirvelu. 2017. “Cloning of HVA22 Homolog from Aloe Vera and Preliminary Study of Transgenic Plant Development.” International Journal of Pure & Applied Bioscience 5(6):1113–21. doi: 10.18782/2320-7051.6033.
Tambaru, Elis. 2015. “Identifikasi Karakter Morfologi Dan Anatomi Stomata Flacourtia Inermis Roxb. Di Kawasan Kampus UNHAS Tamalanrea Makassar.” Jurnal Alam Dan Lingkungan 6(11):37–40.
UPOV. 2015. “Cosmos Cav.” Www.Upov.Int/Edocs/Upov. Retrieved (www.upov.int/edocs/upov).
Wang, Chen-Kuen, Shin-Yun Hsu, Po-Yen Chen, and Kin-Ying To. 2012. “Transformation and Characterization of Transgenic Bidens Pilosa L.” Plant Cell Tiss Organ Cult 109:457–64. doi: 10.1007/s11240-011-0110-9.
Weiss, Julia, Luciana Delgado-Benarroch, and Marcos Egea-Cortines. 2005. “Genetic Control of Floral Size and Proportions.” Int. J. Dev. Biol 49:513–25. doi: 10.1387/ijdb.051998jw.
Yadav, Sheetal, Poojadevi Sharma, Anshu Trivastava, Priti Desai, and Neeta Shrivastava. 2014. “Strain Specific Agrobacterium-Mediated Genetic Transformation of Bacopa Monnieri.” Journal of Genetic Engineering and Biotechnology 12:89–94. doi: 10.1016/j.jgeb.2014.11.003.
Zhang, Xiuren, Rossana Henriques, Shih-Shun Lin, Qi-Wen Niu, and Nam-Hai Chua. 2006. “Agrobacterium-Mediated Transformation of Arabidopsis Thaliana Using the Floral Dip Method.” Nature Protocols 1(2):641–46. doi: 10.1038/nprot.2006.97.
Zhang, Yong-yan, Dong-min Zhang, Yun Zhong, Xiao-jun Chang, Min-lun Hu, and Chun-zhen Cheng. 2017. “A Simple and Efficient in Planta Transformation Method for Pommelo (Citrus Maxima) Using Agrobacterium Tumefaciens.” Scientia Horticulturae 214:174–79. doi: https://doi.org/10.1016/j.scienta.2016.11.033.
Zulkarnain, Rusmadi Rukmana, Hasyimuddin, Masriany, baiq farhatul Wahidah, Nurman, and rahmat fajrin Alir. 2019. “Karakteristik Morfologi Daun Di Kawasan Hutan Hulu Bulu’Ballea, Tinggi Moncong Kabupaten Gowa.” Pp. 48–53 in Prosiding Semnas Biodiversitas Indonesia. UIN Alauddin, Makassar, 3 November 2019. [Indonesia].

Most read articles by the same author(s)

1 2 > >>